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Microchip-Based Purification of DNA from Biological Samples

journal contribution
posted on 2023-05-16, 16:14 authored by Michael BreadmoreMichael Breadmore, Wolfe, KA, Arcibal, IG, Leung, WK, Dickson, D, Giordano, BC, Power, ME, Ferrance, JP, Feldman, SH, Norris, PM, Landers, JP
A microchip solid-phase extraction method for purification of DNA from biological samples, such as blood, is demonstrated. Silica beads were packed into glass microchips and the beads immobilized with sol−gel to provide a stable and reproducible solid phase onto which DNA could be adsorbed. Optimization of the DNA loading conditions established a higher DNA recovery at pH 6.1 than 7.6. This lower pH also allowed for the flow rate to be increased, resulting in a decrease in extraction time from 25 min to less than 15 min. Using this procedure, template genomic DNA from human whole blood was purified on the microchip platform with the only sample preparation being mixing of the blood with load buffer prior to loading on the microchip device. Comparison between the microchip SPE (μchipSPE) procedure and a commercial microcentrifuge method showed comparable amounts of PCR-amplifiable DNA could be isolated from cultures of Salmonella typhimurium. The greatest potential of the μchipSPE device was illustrated by purifying DNA from spores from the vaccine strain of Bacillus anthracis, where eventual integration of SPE, PCR, and separation on a single microdevice could potentially enable complete detection of the infectious agent in less than 30 min.

History

Publication title

Analytical Chemistry

Volume

75

Issue

8

Pagination

1880-1886

ISSN

0003-2700

Department/School

School of Natural Sciences

Publisher

American Chemical Society

Place of publication

Washington

Rights statement

Copyright © 2003 American Chemical Society

Repository Status

  • Restricted

Socio-economic Objectives

Expanding knowledge in the chemical sciences

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